Golgi complex BIOLOGY Faculty of Dentistry 2019 10

Golgi complex BIOLOGY, Faculty of Dentistry 2019. 10. László KŐHIDAI, Ph. D. , Med. Habil. ; Assoc. Prof. Department of Genetics, Cell- and Immunobiology Semmelweis University

Multiphoton fluorescence image of cultured He. La cells

Camillo Golgi (1843 -1926) Nobel prize 1906 "internal reticular apparatus"

Structure ! • Saccules • Tubules • Vesicles structural-functional unit: dictyosome 4 -6 saccles the structure is polarized into sub-compartments cis Golgi medial Golgi cis Golgi network (CGN) trans Golgi network(TGN)

Dictyosomes of the plant cells Synthesis of complex polysaccharides of the cell wall (hemicellulose, pectins)

Relation to the cytoskeleton

Main functions • transport • sorting • transformation • membrane wrapping !

! Golgi – „Traffic manager” of the cell

Main pathways going in and out Golgi !

Cisternal transport d. ER Materials are forwarded together the cisterns Vesicular transport ! Vesicles trasport substances from cystern to cistern

CGN • the peptides arrive from the ER in vesicles • they are N-glycosilated • no sorting in the ER Bidirectional transport of proteins: • soluble, endogeneous proteins of the ER recycled in transport vesicles - retention signal is required • sorting and transport of lysosomal enzymes !

Sorting and modification of lysosomal enzymes Mannose-6 -phosphate (M-6 -P) signaling: • based on the recognition of lysosomal hydrolases • recognition of the “signal patches” (proper 3 D combination of amino acids) is required • main working enzyme: Glc. NAc-phosphotransferase Phosphorylation of the mannoses: • promotes the sorting of these enzymes • prevents the further modifications !

N- and O- glycosylation

Glycosilation in the Golgi Modifications on the N-glycosilation pattern • cis-Golgi: mannose-type oligosaccharides complex oligosaccharides • TGN: substitution with sialic acids - negatively charged O-glycosilation: • takes place mainly in the medial- and trans-Golgi • sidechains of Ser and Thr are glycosilated !

mannse phosphorylation (lysosomal prot. ) Removal of mannose CGN cis-Golgi removal of mannose Labelling w Glc. NAc labelling w galactose Labelling w syalic acid sorting medial-Golgi trans-Golgi TGN !

! Structure of M-6 -P

Significance of M-6 -P labelling !

Other modifications ! • glucose-amino-glycane (GAG) chains • sulphatation (proteoglycanes, Tyr res. of peptides) - TGN • proteolytic modifications - secretion vesicle

Synthesis of lipids in the Golgi glycolipids ceramide sphingomyelin

Main transport pathways from TGN !

Main transport pathways from TGN • endosomal-lysosomal compartment via transport vesicles - M-6 -P receptors • surface membrane - secretion constitutive secretion - transports lipids and peptide components of the surface membrane and the extracellular matrix • exocytosis regulated secretion !

Modifications of secretory vesicles ! • selective aggregation - TGN • further modifications and sorting inactive precursor - active enzyme or hormone (e. g. preproinsulin - insulin) • concentration - loss of water • hydratation - e. g. proteoglygans • uptake some cytoplasmatic substances e. g. histamine

§ Formation of the acrosome Acrosome is a large lysosome and found in sperm § It contains enzymes e. g. hyaluronidase, acid phosphatase… § During the sperm differantiation several small vesicles (primary lysosomes) are formed from Golgi § thesmall vesicles are fused to form a single large lysosome - acrosome

Main pathway going out the Golgi

Alternative pathways • some molecules do not synthesized on the r. ER (e. g. interleukin 1 a and 1 b - IL 1 a-IL 1 b, basic fibroblast growth factor-b. FGF) • these molecules transported by ABC-transporters • other roles of the alternative pathway: - elimination of toxic proteins - regulation of protein concentrations in cytosol !

Defects of sorting mechanism lysosomal enzymes do not enter the late endosomes BUT enzymes enter the constitutive secretory pathway and released I (=inclusion) cell disease: - the M-6 -P signal is not formen on the enzymes - lysosomal enzymes “escape” from the cell - deficient intracellular digestion - the non-digested substances form INCLUSIONS !